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过表达Jagged1促老龄大鼠来源内皮祖细胞增殖和迁移
  • ISSN号:1007-3949
  • 期刊名称:中国动脉硬化杂志
  • 时间:2013
  • 页码:391-396
  • 分类:Q81[生物学—生物工程]
  • 作者机构:[1]成都军区昆明总医院临床实验科 云南省干细胞与组织工程中心,云南省昆明市650032, [2]第三军医大学新桥医院全军心血管疾病研究所,重庆市430037
  • 相关基金:国家自然科学基金(81170316)资助;973基础研究规划项目(2012CB518100)资助
  • 相关项目:Notch1信号调控EPC年龄依赖修复损伤血管作用及机制
中文摘要:

目的探讨过表达Jaggedl对老龄大鼠来源内皮祖细胞增殖和迁移能力的影响。方法PBS冲洗1-2月龄和19~26月龄sD大鼠股骨和胫骨骨髓,Ficoll密度梯度离心法分离单个核细胞,应用含10%FBS的DMEMJF12培养基差速贴壁法进行体外培养,以Dil-ac-LDL与FITC-UEA-1荧光双染和vWF免疫组织化学染色进行鉴定。实验分为四组:对照组(未进行基因转染的老龄大鼠内皮祖细胞组)、PIRES2-EGFP转染组、PIRES2-EGFP-Jaggedl转染组和未转染的年轻大鼠来源内皮祖细胞组。荧光显微镜下计数GFP表达细胞数并计算转染效率;免疫荧光、RT-PCR和Westernblot检测JaggedlmRNA和蛋白表达;Transwell培养和MTT法分别检测细胞迁移和增殖能力。结果免疫组织化学、RT-PCR和Westernblot检测均显示转染后Jaggedl显著增高,PIRES2-EGFP-Jaggedl转染组mRNA和蛋白表达均较对照组显著增强(P〈0.01);与对照组相比,过表达Jaggedl显著增强老龄大鼠来源内皮祖细胞迁移和增殖能力(P〈0.05或P〈0.01)。结论过表达Jaggedl增强老龄大鼠来源内皮祖细胞增殖和迁移能力。

英文摘要:

Aim To investigate the effects of Jaggedl over-expression on aged rat-derived endothelial progenitor cells migration and proliferation. Methods Mononuclear cells were obtained from bone marrow of young ( 1 to 2 month-old) and aged (19 to 26 month-old) Sprague-Dawley rats by Ficoll density gradient centrifugation and cultured with medium DMEM/F12, 48 h later, the suspending cells were translocated to be cultured in new flasks coated with fibronectin, the secondary attached cells were used to perform the further experiments. EPCs were characterized as double posi- tive for Dil-ac-LDL uptake and lectin binding and cells were further identified by vWF expression. The experiments were divided into four groups: control group ( aged rat-derived EPCs without gene transfection), PIRES2-EGFP transfection group, PIRES2-EGFP-Jaggedl transfection group and young rat-derived EPCs group in which transfection was also not performed. The GFP expression cell number was acquired through fluorescence microscope and transfection efficiency was calculated. Immunofluorescence, RT-PCR and Western blot were used to detect the mRNA and protein expression respectively. Cell migration and proliferation were tested by Transwell culture and MTI" assay respectively. Results Immunofluorescence showed that gene transfection markedly promoted Jaggedl expression; RT-PCR and Western blot demon- strated that Jaggedl mRNA and protein expression significantly increased in aged rat-derived EPCs (P〈0.01) compared with control; Over-expression of Jaggedl markedly enhanced aged rat-derived EPCs migration and proliferation compared with control group (P〈0.05 or P〈0.01 respectively). Conclusion Over-expression of Jaggedl accelerates aged rat-derived EPCs migration and proliferation.

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期刊信息
  • 《中国动脉硬化杂志》
  • 北大核心期刊(2011版)
  • 主管单位:中国科学技术协会
  • 主办单位:中国病理生理学会
  • 主编:姜志胜
  • 地址:湖南省衡阳市南华大学内
  • 邮编:421001
  • 邮箱:dmzzbjb@163.net
  • 电话:0734-8160765
  • 国际标准刊号:ISSN:1007-3949
  • 国内统一刊号:ISSN:43-1262/R
  • 邮发代号:42-165
  • 获奖情况:
  • 中国科学技术论文统计源期刊,中国科学引文数据库来源期刊,中国学术期刊综合评价数据库来源期刊
  • 国内外数据库收录:
  • 美国化学文摘(网络版),中国中国科技核心期刊,中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版)
  • 被引量:17016